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A cyclin-dependent kinase family member (PHOA) is required to link developmental fate to environmental conditions in Aspergillus nidulans.

ABSTRACT: We addressed the question of whether Aspergillus nidulans has more than one cyclin-dependent kinase gene and identified such a gene, phoA, encoding two PSTAIRE-containing kinases (PHOAM1 and PHOAM47) that probably result from alternative pre-mRNA splicing. PHOAM47 is 66% identical to Saccharomyces cerevisiae Pho85. The function of this gene was studied using phoA null mutants. It functions in a developmental response to phosphorus-limited growth but has no effect on the regulation of enzymes involved in phosphorus acquisition. Aspergillus nidulans shows both asexual and sexual reproduction involving temporal elaboration of different specific cell types. We demonstrate that developmental decisions in confluent cultures depend upon both the initial phosphorus concentration and the inoculation density and that these factors influence development through phoA. In the most impressive cases, absence of phoA resulted in a switch from asexual to sexual development (at pH 8), or the absence of development altogether (at pH 6). The phenotype of phoA deletion strains appears to be specific for phosphorus limitation. We propose that PHOA functions to help integrate environmental signals with developmental decisions to allow ordered differentiation of specific cell types in A.nidulans under varying growth conditions. The results implicate a putative cyclin-dependent kinase in the control of development.

SUBMITTER: Bussink HJ

PROVIDER: S-EPMC1170733 | biostudies-other | 1998 Jul

REPOSITORIES: biostudies-other

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The PHOA and PHOB cyclin-dependent kinases perform an essential function in Aspergillus nidulans.

Project description:Unlike Pho85 of Saccharomyces cerevisiae, the highly related PHOA cyclin-dependent kinase (CDK) of Aspergillus nidulans plays no role in regulation of enzymes involved in phosphorous acquisition but instead modulates differentiation in response to environmental conditions, including limited phosphorous. Like PHO85, Aspergillus phoA is a nonessential gene. However, we find that expression of dominant-negative PHOA inhibits growth, suggesting it may have an essential but redundant function. Supporting this we have identified another cyclin-dependent kinase, PHOB, which is 77% identical to PHOA. Deletion of phoB causes no phenotype, even under phosphorous-limited growth conditions. To investigate the function of phoA/phoB, double mutants were selected from a cross of strains containing null alleles and by generating a temperature-sensitive allele of phoA in a deltaphoB background. Double-deleted ascospores were able to germinate but had a limited capacity for nuclear division, suggesting a cell cycle defect. Longer germination revealed morphological defects. The temperature-sensitive phoA allele caused both nuclear division and polarity defects at restrictive temperature, which could be complemented by expression of mammalian CDK5. Therefore, an essential function exists in A. nidulans for the Pho85-like kinase pair PHOA and PHOB, which may involve cell cycle control and morphogenesis.

| S-EPMC1462817 | biostudies-other

A Pcl-like cyclin of Aspergillus nidulans is transcriptionally activated by developmental regulators and is involved in sporulation.

Project description:The filamentous fungus Aspergillus nidulans reproduces asexually through the formation of spores on a multicellular aerial structure, called a conidiophore. A key regulator of asexual development is the TFIIIA-type zinc finger containing transcriptional activator Bristle (BRLA). Besides BRLA, the transcription factor ABAA, which is located downstream of BRLA in the developmental regulation cascade, is necessary to direct later gene expression during sporulation. We isolated a new developmental mutant and identified a leaky brlA mutation and the mutated Saccharomyces cerevisiae cyclin homologue pclA, both contributing to the developmental phenotype of the mutant. pclA was found to be 10-fold transcriptionally upregulated during conidiation, and a pclA deletion strain was reduced three- to fivefold in production of conidia. Expression of pclA was strongly induced by ectopic expression of brlA or abaA under conidiation-suppressing conditions, indicating a direct role for brlA and abaA in pclA regulation. PCLA is homologous to yeast Pcl cyclins, which interact with the Pho85 cyclin-dependent kinase. Although interaction with a PSTAIRE kinase was shown in vivo, PCLA function during sporulation was independent of the A. nidulans Pho85 homologue PHOA. Besides the developmental regulation, pclA expression was cell cycle dependent with peak transcript levels in S phase. Our findings suggest a role for PCLA in mediating cell cycle events during late stages of sporulation.

| S-EPMC87069 | biostudies-literature

Functional characterization of a new member of the Cdk9 family in Aspergillus nidulans.

Project description:Cdk9-like kinases in complex with T-type cyclins are essential components of the eukaryotic transcription elongation machinery. The full spectrum of Cdk9/cyclin T targets, as well as the specific consequences of phosphorylations, is still largely undefined. We identify and characterize here a Cdk9 kinase (PtkA) in the filamentous ascomycete Aspergillus nidulans. Deletion of ptkA had a lethal effect in later stages of vegetative growth and completely impeded asexual development. Overexpression of ptkA affected directionality of polarized growth and the initiation of new branching sites. A green fluorescent protein-tagged PtkA version localized inside the nucleus during interphase, supporting a role of PtkA in transcription elongation, as observed in other organisms. We also identified a putative cyclin T homolog, PchA, in the A. nidulans genome and confirmed its interaction with PtkA in vivo. Surprisingly, the Pcl-like cyclin PclA, previously described to be involved in asexual development, was also found to interact with PtkA, indicating a possible role of PtkA in linking transcriptional activity with development and/or morphogenesis in A. nidulans. This is the first report of a Cdk9 kinase interacting with a Pcl-like cyclin, revealing interesting new aspects about the involvement of this Cdk-subfamily in differential gene expression.

| S-EPMC3008270 | biostudies-literature

The role of VosA/VelB-activated developmental gene vadA in Aspergillus nidulans.

Project description:The filamentous fungus Aspergillus nidulans primarily reproduces by forming asexual spores called conidia, the integrity of which is governed by the NF-κB type velvet regulators VosA and VelB. The VosA-VelB hetero-complex regulates the expression of spore-specific structural and regulatory genes during conidiogenesis. Here, we characterize one of the VosA/VelB-activated developmental genes, called vadA, the expression of which in conidia requires activity of both VosA and VelB. VadA (AN5709) is predicted to be a 532-amino acid length fungal-specific protein with a highly conserved domain of unknown function (DUF) at the N-terminus. This DUF was found to be conserved in many Ascomycota and some Glomeromycota species, suggesting a potential evolutionarily conserved function of this domain in fungi. Deletion studies of vadA indicate that VadA is required for proper downregulation of brlA, fksA, and rodA, and for proper expression of tpsA and orlA during sporogenesis. Moreover, vadA null mutant conidia exhibit decreased trehalose content, but increased β(1,3)-glucan levels, lower viability, and reduced tolerance to oxidative stress. We further demonstrate that the vadA null mutant shows increased production of the mycotoxin sterigmatocystin. In summary, VadA is a dual-function novel regulator that controls development and secondary metabolism, and participates in bridging differentiation and viability of newly formed conidia in A. nidulans.

| S-EPMC5421774 | biostudies-literature

A role for NIMA in the nuclear localization of cyclin B in Aspergillus nidulans.

Project description:NIMA promotes entry into mitosis in late G2 by some mechanism that is after activation of the Aspergillus nidulans G2 cyclin-dependent kinase, NIMXCDC2/NIMECyclin B. Here we present two independent lines of evidence which indicate that this mechanism involves control of NIMXCDC2/NIMECyclin B localization. First, we found that NIMECyclin B localized to the nucleus and the nucleus-associated organelle, the spindle pole body, in a NIMA-dependent manner. Analysis of cells from asynchronous cultures, synchronous cultures, and cultures arrested in S or G2 showed that NIMECyclin B was predominantly nuclear during interphase, with maximal nuclear accumulation in late G2. NIMXCDC2 colocalized with NIMECyclin B in G2 cells. Although inactivation of NIMA using either the nimA1 or nimA5 temperature-sensitive mutations blocked cells in G2, NIMXCDC2/NIMECyclin B localization was predominantly cytoplasmic rather than nuclear. Second, we found that nimA interacts genetically with sonA, which is a homologue of the yeast nucleocytoplasmic transporter GLE2/RAE1. Mutations in sonA were identified as allele-specific suppressors of nimA1. The sonA1 suppressor alleviated the nuclear division and NIMECyclin B localization defects of nimA1 cells without markedly increasing NIMXCDC2 or NIMA kinase activity. These results indicate that NIMA promotes the nuclear localization of the NIMXCDC2/ NIMECyclin B complex, by a process involving SONA. This mechanism may be involved in coordinating the functions of NIMXCDC2 and NIMA in the regulation of mitosis.

| S-EPMC2133011 | biostudies-literature

Developmental decisions in Aspergillus nidulans are modulated by Ras activity.

Project description:To better understand how Ras controls development of multicellular organisms, we have chosen Aspergillus nidulans as a model system. When grown on solid medium, this fungus follows a well-defined program of development, sequentially giving rise to several cell types which produce three distinct structures: vegetative hyphae, aerial hyphae, and the conidiophore structure. Here we describe a ras homolog found in this fungus (Aras) and demonstrate that it is an essential gene that regulates the ordered program of development. We created dominant alleles of this gene and expressed them to different levels in order to vary the ratio of GTP-bound (active) to GDP-bound (inactive) A-Ras protein. When the amount of active Ras is large, nuclear division proceeds, but further development is inhibited at the early step of germ tube formation. At an intermediate level of active Ras, aerial hypha formation is inhibited, while at a low level, conidiophore formation is inhibited. Maintenance of an even lower level of the active Ras is essential for initiation and progression of conidiophore formation, the final stage of development. When the level of active Ras is artificially lowered, each stage of development is initiated prematurely except germination, the initial stage of development. Therefore, the progression of the ordered developmental pathway of A. nidulans is dependent upon an initial high level of active Ras followed by its gradual decrease. We propose that several concentration threshold exist, each of which allows development to proceed to a certain point, producing the proper cell type while inhibiting further development.

| S-EPMC359053 | biostudies-other

Identification of developmental regulatory genes in Aspergillus nidulans by overexpression.

Project description:Overexpression of several Aspergillus nidulans developmental regulatory genes has been shown to cause growth inhibition and development at inappropriate times. We set out to identify previously unknown developmental regulators by constructing a nutritionally inducible A. nidulans expression library containing small, random genomic DNA fragments inserted next to the alcA promoter [alcA(p)] in an A. nidulans transformation vector. Among 20,000 transformants containing random alcA(p) genomic DNA fusion constructs, we identified 66 distinct mutant strains in which alcA(p) induction resulted in growth inhibition as well as causing other detectable phenotypic changes. These growth inhibited mutants were divided into 52 FIG (Forced expression Inhibition of Growth) and 14 FAB (Forced expression Activation of brlA) mutants based on whether or not alcA(p) induction resulted in accumulation of mRNA for the developmental regulatory gene brlA. In four FAB mutants, alcA(p) induction not only activated brlA expression but also caused hyphae to differentiate into reduced conidiophores that produced viable spores from the tips as is observed after alcA(p)::brlA induction. Sequence analyses of the DNA fragments under alcA(p) control in three of these four sporulating strains showed that in two cases developmental activation resulted from overexpression of previously uncharacterized genes, whereas in the third strain, the alcA(p) was fused to brlA. The potential uses for this strategy in identifying genes whose overexpression results in specific phenotypic changes like developmental induction are discussed.

| S-EPMC1206365 | biostudies-other

International Space Station conditions alter genomics, proteomics, and metabolomics in Aspergillus nidulans.

Project description:The first global genomic, proteomic, and secondary metabolomic characterization of the filamentous fungus Aspergillus nidulans following growth onboard the International Space Station (ISS) is reported. The investigation included the A. nidulans wild-type and three mutant strains, two of which were genetically engineered to enhance secondary metabolite production. Whole genome sequencing revealed that ISS conditions altered the A. nidulans genome in specific regions. In strain CW12001, which features overexpression of the secondary metabolite global regulator laeA, ISS conditions induced the loss of the laeA stop codon. Differential expression of proteins involved in stress response, carbohydrate metabolic processes, and secondary metabolite biosynthesis was also observed. ISS conditions significantly decreased prenyl xanthone production in the wild-type strain and increased asperthecin production in LO1362 and CW12001, which are deficient in a major DNA repair mechanism. These data provide valuable insights into the adaptation mechanism of A. nidulans to spacecraft environments.

| S-EPMC9098231 | biostudies-literature

Gene expression in Aspergillus nidulans at different developmental stages

Project description:Microarray analysis was used to compare different developmental stages of the filamentous fungus Aspergillus nidulans. Cells were grown for various times and comparisons made between: 1. Isolated conidia (0 hours) and isotropically expanding cells (3 hours). 2. Isotropically expanding cells (3 hours) and polarly extending cells (5 hours). 3. Undifferentiated vegetative hyphae (18 hours) and synchronously conidiating cultures (42 hours). Keywords: Developmental stage comparison

2008-05-01 | GSE7698 | GEO

Elucidation of functional markers from Aspergillus nidulans developmental regulator FlbB and their phylogenetic distribution.

Project description:Aspergillus nidulans is a filamentous fungus widely used as a model for biotechnological and clinical research. It is also used as a platform for the study of basic eukaryotic developmental processes. Previous studies identified and partially characterized a set of proteins controlling cellular transformations in this ascomycete. Among these proteins, the bZip type transcription factor FlbB is a key regulator of reproduction, stress responses and cell-death. Our aim here was the prediction, through various bioinformatic methods, of key functional residues and motifs within FlbB in order to inform the design of future laboratory experiments and further the understanding of the molecular mechanisms that control fungal development. A dataset of FlbB orthologs and those of its key interaction partner FlbE was assembled from 40 members of the Pezizomycotina. Unique features were identified in each of the three structural domains of FlbB. The N-terminal region encoded a bZip transcription factor domain with a novel histidine-containing DNA binding motif while the dimerization determinants exhibited two distinct profiles that segregated by class. The C-terminal region of FlbB showed high similarity with the AP-1 family of stress response regulators but with variable patterns of conserved cysteines that segregated by class and order. Motif conservation analysis revealed that nine FlbB orthologs belonging to the Eurotiales order contained a motif in the central region that could mediate interaction with FlbE. The key residues and motifs identified here provide a basis for the design of follow-up experimental investigations. Additionally, the presence or absence of these residues and motifs among the FlbB orthologs could help explain the differences in the developmental programs among fungal species as well as define putative complementation groups that could serve to extend known functional characterizations to other species.

| S-EPMC3053368 | biostudies-literature

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